1033 replies to this topic

[QuestforLife]

Just wanted to post a quick link to another thread someone started on skin rejuvenation. One of the chemicals identified is a ROCK inhibitor, and a quick search shows statins have been used topically for improved wound healing.

https://www.longecit...on/#entry872303

So I think it's worth having a go at creating a topical statin solution. At the moment i only have atorvastatin, which is a little heavy for skin penetration, but is lipophillic, so should dissolve well in lots of skin creams.

 

So I made a statin skin cream. I used Atopalm for the base as it has chloresterol in it, which should counter the negative effects of statins making skin scaly.  I also used Atopalm because it has been shown to reduce systemic inflammation when used on old skin, and is also proven to penetrate the stratum corneum. 

 

See: https://onlinelibrar....1111/jdv.15540

 

Background: While increased levels of circulating inflammatory cytokines in chronologically

aged humans have been linked to the development of aging-associated chronic disorders (e.g.,
cardiovascular disease, type II diabetes, osteoporosis and Alzheimer’s disease), approaches that
reduce circulating cytokines are not yet available. In chronologically aged mice, we recently
demonstrated that epidermal dysfunction largely accounts for age-associated elevations in
circulating cytokine levels, and that improving epidermal function reduced circulating cytokine
levels.

 

Objective: We performed a pilot study to determine whether improving epidermal
function reduces circulating proinflammatory cytokine levels in aged humans. Methods: Thirtythree aged humans were treated twice-daily for 30 days, with ≈3 ml of an emollient, previously
shown to improve epidermal function, while untreated, aged humans and a cohort of young
volunteers served as controls. Changes in epidermal function and levels of three key, age-related,
plasma cytokines (IL-1β, IL-6 and TNFα) were measured at baseline and after treatment, using
Luminex 200™ system. Results: We also found significantly higher baseline levels of IL-1β, IL6 and TNFα in aged vs. young humans (p<0.001), as previously reported. Topical applications of
the barrier repair emollient significantly enhanced epidermal permeability barrier function
(p<0.01) and stratum corneum hydration (p<0.05). In parallel, circulating levels of IL-1β and IL6 normalized, while TNFα levels declined substantially.

 

Conclusion: The results of this
preliminary study suggest that a larger clinical trial should be performed to confirm whether
improving epidermal function also can reduce circulating proinflammatory cytokine levels in
aged humans, while also possibly attenuating the downstream development of chronic
inflammatory disorders in the aged humans.

 

It's total guesswork but I've mixed in 100mg of simvastatin and 60mg of telmisartan with 40ml of Atopalm, which I'm applying to the face only in the morning. Simvastatin and telmisartan dissolved very well in the Atopalm. It's hard to say exactly how much I'm using a day but I'd estimate I'll use up the skin cream in 2 months, which means my skin is getting about 1.7mg of statins a day (maybe too much?).

 

I've been using it for a week. No side effects to report. Skin appearance is noticeably improved; the Atopalm seems to remain in the skin all day markedly improving moisture content. At this point it is impossible to attribute any effects to my additional ingredients. 

 

My future plan is to also incorporate azithromycin into the skin cream, as a way of destroying senescent skin cells and making room for any progenitors the statin+sartan combination have created. Unfortunately azithromycin is a heavy molecule, so skin penetration may be limited.

[QuestforLife]

After 10 days of treatment once per day with my statin cream, results were not as expected (other than the generally good moisturizing effect of the Atopalm), so I added 500mg of Azithromycin with another 10ml of Atopalm base to the mixture.

Theory is the Azithromycin will kill any senescent cells in the skin and the apoptosis products will then encourage proliferation and replacement by any progenitors the statins have created.

I'll try this for 10 days and report back.

[Andey]

I ve just finished my streak on this protocol/method.

Atorvastatin 10mg every other day and 12.5 mg of losartan everyday for two month.

So far nothing to report, but I plan to do some basic lab tests at the end of the month. Ive done one in the process and the only obvious thing was a my cholesterol down like 20%.

 

Plan to repeat it after two month or so.

[QuestforLife]

I ve just finished my streak on this protocol/method.
Atorvastatin 10mg every other day and 12.5 mg of losartan everyday for two month.
So far nothing to report, but I plan to do some basic lab tests at the end of the month. Ive done one in the process and the only obvious thing was a my cholesterol down like 20%.

Plan to repeat it after two month or so.

I'm not sure what you should expect to see; the protocol was designed for middle aged men who had no symptoms, but nevertheless had early signs of impaired arterial health as measured by pulse wave velocity and flow mediated dilation. One month treatment was sufficient to correct this, with the benefits not declining to baseline till 5 months after ceasing treatment.

I think you should probably try a shorter protocol next time, as 20% reduction in cholesterol shows you have edged above the 'pleiotropic range' even though you're only dosing the statin every other day. Or perhaps atorvastatin is too strong even at that dose.

I plan to do 3 treatments a year, with my next one in late July/early August. I've got some more ideas about combining it with a more powerful telomerase activator, but I won't post on that yet till I have it clearer in my head.

[Andey]

I think you should probably try a shorter protocol next time, as 20% reduction in cholesterol shows you have edged above the 'pleiotropic range' even though you're only dosing the statin every other day. Or perhaps atorvastatin is too strong even at that dose.

 

   I think my dosage/response is pretty much in line with what should be expected from low dose statin, you could check it in studies. (BTW with your dosages it should be even higher)

Edited by Andey, 15 June 2019 - 07:04 PM.

[aribadabar]

Most of the testamonials on Longecity are n=1. In any case my theory about statins reducing epigenetic age comes from knowledge of the action of ROCK inhbitors, and the methylation age of several others on statins. But yes, evidence is very tentative at this time.

 

I also have wondered how robust this 2.4-year de-aging is. Is it within the normal scatter or outside the noise and therefore a veritable outlier?

Do Zymo give any details of the precision of their test result?

[QuestforLife]

I think my dosage/response is pretty much in line with what should be expected from low dose statin, you could check it in studies. (BTW with your dosages it should be even higher)

I only say that because I'm sure one of the Janic studies said the low dose protocol didn't effect cholesterol.

I've tried various dosing schedules. When I did it for a month it was at 10mg/day atorvastatin. When I took it at a higher dose it was only for 2 weeks.

Edited by QuestforLife, 16 June 2019 - 09:04 AM.

[QuestforLife]

I also have wondered how robust this 2.4-year de-aging is. Is it within the normal scatter or outside the noise and therefore a veritable outlier?
Do Zymo give any details of the precision of their test result?

This is what I received from Zymo, when I asked about the reproducibility of their blood test:

Average
SD

Technical

same blood sample that analyze independently

0.9899

Biological

multiple blood samples that collected from same individuals at the same time

1.4224

Which means if they retested my same blood sample a bunch of times the scatter of results would give a standard deviation of about 1 year.

Edited by QuestforLife, 16 June 2019 - 09:12 AM.

[Andey]

I only say that because I'm sure one of the Janic studies said the low dose protocol didn't effect cholesterol.

I've tried various dosing schedules. When I did it for a month it was at 10mg/day atorvastatin. When I took it at a higher dose it was only for 2 weeks.

 

 

  Ive looked at Janic studies, there is no mention of it and in one of those LDL-C was lowered little below 20% too.

Having said that I think it would be sensible to go next time for a short half life statin like fluvastatin that Janik have used. Unfortunately they are hard to come by as all modern statins are long lasting therefore more effective.

[QuestforLife]

  Ive looked at Janic studies, there is no mention of it and in one of those LDL-C was lowered little below 20% too.

Having said that I think it would be sensible to go next time for a short half life statin like fluvastatin that Janik have used. Unfortunately they are hard to come by as all modern statins are long lasting therefore more effective.

 

Which paper showed the 20% reduction?

 

'Long‑term improvement of arterial wall characteristics in patients with diabetes mellitus type 1 using cyclic, intermittent treatment with a low‑dose fluvastatin and valsartan combination' from 2015, shows no change in either total or LDL-cholesterol with one month of 10mg fluvastatin daily. It's a good paper and I recommend you read it if you haven't, as it shows you can repeat the treatment and regain the benefits.

 

I've also looked to acquire fluvastatin and been unable to do so. It shouldn't matter as supposedly the arterial benefits of this protocol are independent cholesterol reduction, but I would prefer to keep my cholesterol levels at what I believe to be a (slightly higher than is normally considered) optimal  range.

[Andey]

Which paper showed the 20% reduction?

 

'Long‑term improvement of arterial wall characteristics in patients with diabetes mellitus type 1 using cyclic, intermittent treatment with a low‑dose fluvastatin and valsartan combination' from 2015, shows no change in either total or LDL-cholesterol with one month of 10mg fluvastatin daily. It's a good paper and I recommend you read it if you haven't, as it shows you can repeat the treatment and regain the benefits.

 

I've also looked to acquire fluvastatin and been unable to do so. It shouldn't matter as supposedly the arterial benefits of this protocol are independent cholesterol reduction, but I would prefer to keep my cholesterol levels at what I believe to be a (slightly higher than is normally considered) optimal  range.

Thanks, I havent read that, probably because they used different spelling for Janic and pubmed doesnt group it as one author

 

 

Ive seen a reduction in

The effects of low-dose fluvastatin and valsartan combination on arterial function:
https://sci-hub.se/h...pubmed/22385885

Here LDL-C was lowered from 3.7 to 3.1

Same fluvastatin 10mg 

Edited by Andey, 17 June 2019 - 09:19 AM.

[QuestforLife]

Thanks, I havent read that, probably because they used different spelling for Janic and pubmed doesnt group it as one author

 

 

Ive seen a reduction in

The effects of low-dose fluvastatin and valsartan combination on arterial function:
https://sci-hub.se/h...pubmed/22385885

Here LDL-C was lowered from 3.7 to 3.1

Same fluvastatin 10mg 

 

That's interesting. But the reduction only has a P-value of 0.14 compared to the control group, so not significant. Still, it probably is real. 

 

Personally, I'll just eat some more bacon!  

[Andey]

BTW I noticed that my skin looks a tad better after the cycle. Its hard to judge it on the face but on the body skin looks better hydrated

[QuestforLife]

BTW I noticed that my skin looks a tad better after the cycle. Its hard to judge it on the face but on the body skin looks better hydrated

I experienced the same thing, I think I reported it upthread. I only noticed it from a photo and seeing how the skin on my cheek was much less dry than in a photo a few months prior.

[QuestforLife]

Telomerase Activators Revisited
I and others have posted some of this elsewhere, but in the interests of clarity and ease of reference, I thought it would bear repeating and expanding on the (often suspected) molecular structure of various small molecule telomerase activators.
I also have an interest in finding the best telomerase activator to go along with my statin-sartan protocol, which I will elaborate on later.
Cycloastragenol and TA-65
Cycloastragenol (CA) has a molecular weight of 490g/M and the following molecular structure:

The many OH groups possibly explain it’s known poor bioavailability.
Bill Andrews has stated in several interviews [for example recently here, https://www.youtube....?v=Tx9yG6iTROQ]that he does not believe, contrary to general perception, that TA-65 is cycloastragenol due to the fact Sierra Sciences have found telomerase activation from the former but not the latter. 
Looking at the most recent TA Sciences patent [https://patents.goog...rley&sort=new],cycloastragenol is prominently mentioned, and I find it hard to  believe that this is pure misdirection. They do however allude to other substances, such as C3-(L)-Valyl-cycloastragenol, which has a heavier molecular weight of 625, but at least 10x the bioavailability (at least in dogs) compared to CA – probably largely due to the removal of OH groups (rather like how pterostilbene is heavier than resveratrol but with a much longer half life due to the replacement of two of the three OH groups with CH3). If I’m right, then TA-65 looks like this:

Experiments in vivo have shown TA-65 is far more bioavailable than cycloastragenol, although noting this data came from TA Sciences, [https://m2j3j7n4.sta...id=4ec51f66cb].
This has been suggested to be due to improved formulation of the capsule, but perhaps it is actually because they are different molecules.
TAM-818
Supposedly a far more potent telomerase activator is Bill Andrews’ TAM818. He claims in vitro that TAM818 can produce 16% [https://www.youtube....?v=Tx9yG6iTROQ]of the telomerase produced by a cancerous HELA cell (presumably 100% would allow unlimited proliferation and potential immortality). Looking at the relevant Sierra Science’s patent [https://patents.google.com/patent/US20090143451A1/en?inventor=William+H.+Andrews&sort=new], it is a isoxazole molecule, most likely this:
 
Interestingly the ring structure is a common feature with the CA-like molecules used by TA Sciences, but TAM818 is much smaller, with a MW of only 234 and no pesky OH groups to be quickly digested by the liver – so although there is no actual bioavailability data on TAM818, there are reasons to be optimistic it will be well absorbed. In fact in a personal communication with me, Bill Andrews stated that this molecule was designed according to Lipinkski’s Rule of 5 [https://en.wikipedia..._rule_of_five],i.e. with bioavailability in mind.
Other telomerase activators
One such is Product B from Isagenix, stated by Bill Andrews to be a telomerase activator [https://www.youtube....v=eyEsGzv2r_k].Unfortunately it has a long list of natural ingredients, and it is not clear which ones are the active components, or in what combination or concentration they are required. Therefore it will not be discussed further here.
Telobooster is the invention of a Frenchman, Richard Caterini – it is not remarkable in that it is just cycloastragenol and astralogosides IV mixed with some other things (selenium, Vit C, Vit E, TMG, Lysine and Folate), but it does have data in humans over a number of years – and it seems superior to TA-65, at least from the data on leukocyte telomere length [http://www.bluepenjo...er/Caterini.pdf , http://www.bluepenjo...y/Caterini.pdf, http://telobooster.n..._05_12-Eng.pdf, https://patents.goog...R3016290A1/en].
More intriguing is the idea of synergies between natural molecules, which individually only have a very small effect on telomerase, but together do something substantial. For example Telomere Plus is discussed in the following paper: [https://www.research...ing_Compounds].The best blend consisted of broccoli extract, Rhodolia, astralagus extract and Vitamin D, and in vitro at least, was better than TA-65. It is intriguing that the components are all reasonably bioavailable, and the synergy is highly interesting – especially the combination of broccoli seed extract (a NRF2 activator and HDAC inhibitor) with astralagus. I wonder if the combination with CA or TA-65 would also be synergistic?
Why am I revisiting this?
Even Bill Andrews, the premier researcher in the field of small molecule telomerase activators, is downbeat when it comes to the ability of any currently available small molecules to produce sufficient telomerase to retard aging, let along reverse it. TAM818 in vitro is 16% of the way there, but who knows what it does in vivo, and everything else is known to be much weaker. That is why Bill has become involved in gene therapy.
But I think there might still be hope. The reason I say this is based on the amazing phenomenon of conditional reprogramming  I’ve discussed this at length up-thread. This is when mortal somatic human cells are put in the right kind of culture to allow them to proliferate INDEFINITELY. How do they do this? Well the key seems to be that the cells are transformed back into progenitor cells for the duration of the treatment, differentiating back into  somatic cells when treatment is withdrawn (with no risk of cancer). The intriguing part for us is that there are TWO mechanisms that work together. Neither is sufficient alone.
One is telomerase activation. This was originally done accidentally by ‘feeding’ the somatic cells with the products of senescent mouse embryonic cells. These feeder cells were radiated to stop them overgrowing the somatic cells, but it turns out they still produce telomerase, and quite a lot of it. In later experiments they discovered the telomerase is somehow contained in the apoptosis products of these cells (could we use senolytics to get the same effect?). And in even more recent experiments they have discovered that these feeder cells aren’t needed if you deliver an alternatively spliced form of P53, called delta133p53alpha. At the moment this is done as a simple form of gene therapy, but perhaps in the future small molecules might be a possibility.
The second necessary factor is a Rho-kinase inhibitor, or ROCK inhibitor for short. These chemicals disrupt the cytoskeleton of cells, allowing them to return to an adult stem like state. This also increases telomerase, but much less so than the first mechanism discussed above. Specific ROCK inhibitors aren’t widely available (Fasidil, Y-27632), but statins are also known to be ROCK inhibitors. I’ve posted to death (up thread) on Janic et al. and his work rejuvenating arteries using a short term, low dose, statin and sartan protocol, and this I believe is the mechanism behind his success. Telomerase is also increased by the protocol (hence it’s inclusion in my thread), and interestingly statin users are known to have longer telomeres than controls and just might even have a younger epigenetic age as well (see my theory in the members area), as you’d expect from de-differentiation.
But the real point of me discussing this is the fact that even the large amount of telomerase produced by feeder cells was not sufficient to allow these cells to proliferate indefinitely. In the Bill Andrew’s school of thought this means that telomerase activation was insufficient (less than 100% Hela). But the addition of the ROCK inhibitor only adds a little telomerase to the total but does permit indefinite proliferation. Plus the delta133p53alpha generated telomerase is less than that released by feeder cells, but the addition of a ROCK inhibitor STILL allowed unlimited proliferation, even though the total telomerase was less. Added to this, the ROCK inhibitor alone caused longer proliferation than delta133p53alpha alone, showing that extended proliferation isn’t just about the quantity of telomerase.
So, in that incredibly roundabout way that brings me to my conclusion. TAM818 might only be 16% of HELA and insufficient ALONE to permit unlimited proliferation of somatic cells. But what about in combination with a ROCK inhibitor? The statin-sartan protocol already increases telomerase 200-300% in vivo (https://www.liebertp...9/rej.2015.1722). What more might be possible if we add TAM818 to these progenitor cells? Might we be on the verge of a true antiaging breakthrough? Or is the telomerase activation available through small molecules like TAM818 less than that provided by the statin-sartan combination and hence add nothing? There is only one way to find out…

[QuestforLife]

Another possible telomerase activator is the weak androgen, Danazol. Molecular weight 337, structure:

 

Again note the structural similarity to other known telomerase activators (rings, and also Oxygen and Nitrogen substituted into one ring as with TAM-818).

 

There is evidence for its efficacy at a high dose (800mg/day) in those with telomere diseases in a Phase I-II trial, see: https://www.nejm.org...6/NEJMoa1515319

 

And a Phase II trial at 200mg/day and 400mg/day is currently underway, see here: https://clinicaltria...udy/NCT03312400

 

Lower dose is probably to avoid side effects, of which the most concerning is potential liver damage. 

 

Edited by QuestforLife, 20 June 2019 - 04:25 PM.

[QuestforLife]

Another telomerase activator is Epitalon (thanks to aribadabar for the tip).  Epitalon has a MW of 390 and the following structure:

Epitalon is simply 4 amino acids put together (a tetra-peptide): alanine, glutamic acid, aspartic acid and glycine. It can’t survive digestion so must be injected – normally subcutaneously with an insulin needle after being combined with bacteriostatic water, most often at 5mg once or twice a day. Obviously the main concern with this is finding a reliable, safe source.
Epitalon has only been researched so far in Russia. I could find the following paper: https://www.ncbi.nlm...pubmed/12937682 (available on sci-hub), which showed that after 4 days of 0.05ug/ml epitalon (achievable at 5mg/day injected), in vitro human fibroblasts had telomerase levels comparable or even higher than telomerase positive HELA cells! Another paper for which I can only see the abstract: https://www.ncbi.nlm...ubmed/15455129, claimed fibroblasts treated in this way extended their proliferation from 34 to 44 passages.
Common dosing from those online is 20 consecutive days once or twice a year, so this would be ideal for combining with the statin-sartan protocol. There are anecdotal reports of epitalon being effective at elongating human telomeres in vivo, see: https://www.reddit.c..._with_epitalon/

Edited by QuestforLife, 21 June 2019 - 10:29 AM.

[QuestforLife]

So I made a statin skin cream. I used Atopalm for the base as it has chloresterol in it, which should counter the negative effects of statins making skin scaly.  I also used Atopalm because it has been shown to reduce systemic inflammation when used on old skin, and is also proven to penetrate the stratum corneum. 

 

See: https://onlinelibrar....1111/jdv.15540

 

It's total guesswork but I've mixed in 100mg of simvastatin and 60mg of telmisartan with 40ml of Atopalm, which I'm applying to the face only in the morning. Simvastatin and telmisartan dissolved very well in the Atopalm. It's hard to say exactly how much I'm using a day but I'd estimate I'll use up the skin cream in 2 months, which means my skin is getting about 1.7mg of statins a day (maybe too much?).

 

I've been using it for a week. No side effects to report. Skin appearance is noticeably improved; the Atopalm seems to remain in the skin all day markedly improving moisture content. At this point it is impossible to attribute any effects to my additional ingredients. 

 

My future plan is to also incorporate azithromycin into the skin cream, as a way of destroying senescent skin cells and making room for any progenitors the statin+sartan combination have created. Unfortunately azithromycin is a heavy molecule, so skin penetration may be limited.

 

I've almost finished my home made skin cream. My skin DOES look better, but this may simply be down to the excellent moisturising properties of the Atopalm, although the skin still looks improved even after a thorough face wash. Once I've completely finished the home made skin cream I plan to conduct a positive control by using Atopalm on its own for a month and compare the results. This may have to wait for a month however, to avoid confusing the results with my next statin-sartan protocol iteration (begun yesterday).

[QuestforLife]

A note on pioglitazone

 

For a while I’ve been wondering whether pioglitazone would be a useful addition to my statin-sartan protocol. We know statins and sartans are synergistic in improving the health of the circulatory system – I expect this is due to the simultaneous intracellular effect on both the cytoskeleton (statin) and the angiotensin system (sartan). We also know from a separate source(http://www.jbc.org/c...jbc.M114.624841)  that pioglitazone in addition to a ROCK inhibitor increases the colony forming efficiency of iPSCs by 2-3x compared to a ROCK inhibitor alone. Pioglitazone is a PPARy activator, and it is believed that this increases the expression of angiotensin converting enzyme (ACE). It may have more mechanisms but at this time it appears to be acting in much the same way as a Sartan would, and therefore need not be added to the protocol.

 

On the other hand, pioglitazone is known to be able to stimulate new adipocyte creation, which would certainly be useful for looking younger, so I'll keep researching.

 

Next iteration of the protocol

 

In the meantime I've started my latest iteration of the statin-sartan protcol:

• atorvastatin 5mg/day for 30 days (reduced from 10mg/day due to long half life compared with fluvastatin)
• losartan 25mg/day for 30 days (I may reduce this depending on effects on blood pressure, which I want to avoid)
• epitalon subq 5mg/day for 20 days (added for telomerase activation)

Other than these I will continue my normal lifestyle and minimalist stack: intermittent fasting, whey on weights days, NAC on non weight days, also Glycine, B6 and Magnesium Citrate daily.

Edited by QuestforLife, 04 July 2019 - 09:26 AM.

[QuestforLife]

 

Next iteration of the protocol

 

In the meantime I've started my latest iteration of the statin-sartan protcol:

• atorvastatin 5mg/day for 30 days (reduced from 10mg/day due to long half life compared with fluvastatin)
• losartan 25mg/day for 30 days (I may reduce this depending on effects on blood pressure, which I want to avoid)
• epitalon subq 5mg/day for 20 days (added for telomerase activation)

Other than these I will continue my normal lifestyle and minimalist stack: intermittent fasting, whey on weights days, NAC on non weight days, also Glycine, B6 and Magnesium Citrate daily.

 

Latest iteration of the protocol is complete.

 

Variations from the plan: upped epitalon at the half-way mark to 10mg/day
Tried both 5mg/day and 10mg/day of atorvastatin; felt better on 5mg/day

I ceased atorvastatin and losartan on day 20 (when the epitalon ran out)

 

Some observations: I tentatively attribute these to the addition of the epitalon

• First few days I felt unusually tired, but this passed.
• I experienced slightly watering eyes after the first few epitalon injections, but this also passed.
• I experienced worse than usual hayfever throughout the protocol (it is hayfever season).
• I experienced an insect bite that swelled up much more than I have experienced before during the protocol; again it is impossible to say this is definitely due to the epitalon.
• I have made very good progress in my weights training during the protocol period.
• My skin looks improved, possibly more so than I experienced from the statin-sartan protocol without epitalon.
• It is possible that my hair is growing faster.

I will repeat the protocol in the autumn and then take a Life Length telomere test.

Edited by QuestforLife, 22 July 2019 - 08:37 AM.

[QuestforLife]

Another benefit I've noticed since I've finished the last protocol is the resolution of a split big toe nail that occurred due to a biotin deficiency when I was on Keto. 3 months on a normal
diet had not resolved the problem, but the single course with epitalon has almost completely healed the damage.

I now intend to take a break and then try out endoluten, the oral form of epithalamin.

I've also discovered another researcher, who has independently been investigating ROCK inhibitors as part of an in Vivo rejuvenation effort. His summary is well worth reading:

https://www.academia...enation_in_situ

[QuestforLife]

I thought I'd write this post to explain a little bit more about my view of aging.

I look at aging in a very simple way. Cells are like little powerplants that make their own energy and manufacture the molecules they need. They also clean up the waste this produces, which includes the molecules they've made. But like any waste disposal process, it isn't perfect. Hence to ensure continual (immortal) health a cell has to divide to keep levels of metabolic waste from reaching critical levels.

This ability of dividing cells is one reason bacteria generally don't age, children are healthier than adults, why some animals that stay in a neonatal (child-like)state (i.e. my avatar the axolotl) appear not to age, why animals that grow all their lives appear not to age, and why aging seems to start as soon as growth ceases and adulthood starts.

If cell division ceases then the cell has only cell cleaning to keep it healthy, and this is where autophagy comes in. As soon as autophagy gets behind we get cellular growth (rather than division) and hyperfunction (followed inevitably by dysfunction). Old cells get bigger.

It is interesting that the smaller a cell is, the more stem-like. Pluripotent or embryonic cells are the smallest. Among mesenchymal stem stems, the smaller mesenchymal stem cells are the most totipotent.

So let's talk about telomeres. Telomeres are what is needed to keep cell division going, and to keep harmful metabolic aggregates below a certain level, and to keep cells small. In cells that don't divide by design, or divide very slowly (neurons and cardiomyocyte cells come to mind) there are always other dividing cells on which those non dividing cells depend (glial and astrocytes for neurons, endothelial cells for cardiomyocytes, satellite cells for the muscle, etc.) and which in some cases (glial and astrocytes for neurons) are actually responsible for helping clear the waste (amyloid beta and tau for example).

Additionally, gene expression changes as telomeres shorten,and not only does this slow cell division, but it also slows all protein turnover within the cell. Hence autophagy is failing as a direct result of telomere loss in dividing cells, in addition to the waste disposal problem downstream in some non-dividing cells.

So as we age the cells of the body are getting less numerous, bigger, and less stem-like (and this is probably where methylation changes come in).

Now stem cells spawn progenitor cells, and progenitor cells do much of the dividing that produces somatic cell lines. We get more division in tissues that need more replacement, i.e. more cell division in blood than muscle, for example, so telomeres are shorter in blood than muscle. But telomere rate of loss is about the same for all tissues as we age (https://www.nature.c...cles/ncomms2602). Which tells us that there are less progenitor cells in say muscle than blood (less cells divide less often equals same loss). Hence tissue maintenance is failing across the whole body with age as a result of shortening telomeres, and also leads to a metabolic waste problem in the cells that remain.

So what can we do as a present day intervention?

First things first. As we get older autophagy upregulators are a necessity (some combination of glucosamine, coffee, rapamycin,fasting, exercise, etc.) to keep cells smaller and cleaner until such as time as the telomere problem is addressed.

Secondly, telomeres. We need powerful telomerase activators. This thread has gone some way towards addressing this.

Thirdly, loss of stemness and epigenetic aging. This thread goes some way to addressing this by shrinking cells using ROCK inhibitors and this also contributes their proliferative ability.

I also have a senolytic/anti cancer protocol in preparation(https://www.ncbi.nlm...les/PMC6520007/) which I've mentioned elsewhere, using liposomal Vit C, and two synergistic antibiotics. I will post on this when I have tried it. Another member will also be trialling this. Senolytics are useful in that they can clear out death resistant cells that have become permanently senescent and can't be rejuvenated by the above methods, and also reverse some of the downstream harmful metabolic effects we talked about. But I wouldn't use them as a monotherapy due to the increased requirement for more cell division, which might exacerbate the problem in the long run.

[QuestforLife]

I call this the 'Cell Size Theory of Aging' as it neatly captures cellular hyperfunction and metabolic aging, cellular senescence, telomere shortening and stem cell aging as well as methylation aging.

[QuestforLife]

So it turns out both the benefits and side effects of statins are due to blocking differentiation, at least according to this paper:

 

https://www.physiolo...cell.00406.2014

 

More interesting is a detailed look at the heterogeneous lot that is mesenchymal stem cells:

 

https://journals.plo...al.pone.0120137

 

From the abstract:

 

 Importantly, we observed that simvastatin increased the percentage of a subset of smaller MSC, which also were actively proliferating. The association of MSC decreased size with increased pluripotency and the accumulating evidence that statins may prevent cellular senescence led us to hypothesize that simvastatin induces a smaller subpopulation that may have increased ability to maintain the entire pool of MSC and also to protect them from cellular senescence induced by long-term cultures/passages in vitro. These results may be important to better understand the pleiotropic effects of statins and its effects on the biology of cells with regenerative potential.

 

In the paper they didn't know whether the statin was selectively allowing for the expansion of smaller stem cells already present in the pool, whilst inhibiting the larger cells, or whether larger cells were being turned into smaller cells by the statin. Excitingly, we know from the in vitro work with ROCK inhbitors, that it is the latter.

 

So it is not just terminally differentiated somatic cells like endothelial or keratinocytes that can be de-differentiated by statins, but also stem cells too. So my statin-sartan protocol should have benefits all the way down the tree to the roots that are the stem cell pools.

 

These papers also reinforce why this protocol should be intermittent - as differentiation is required by the body constantly to replace lost somatic cells, and reducing this for too long will obviously not be beneficial. 

 

My recommended cycle is 20 days with 5mg atrovastatin, 12.5mg losartan and a telomerase activator.

[QuestforLife]

One other note:

 

Whilst I am off the statin-sartan oral cycle, I am re-trying my statin skin cream but with some changes.

 

• I have approximately doubled the concentration (200mg rather than 100mg of simvastatin in 50ml atopalm cream)
• I have removed the telmisartan and azithromycin
• I am only applying it twice a week but immediately after dermarolling

The reason I'm trying dermarolling is that there is good evidence statin skin creams help heal skin injuries (ref upthread), so maybe skin rejuvenation requires better penetration of the stratum corneum and potentially also inflammatory growth signalling. The punctures from derma needling should provide both.

[QuestforLife]

Some more ideas on aging and skin aging in particular: Stem cells are activated by stressors. Over a lifetime it makes sense that the more easily activated stem cells (and there is a range of responsiveness in all cells) will be depleted first – because asymmetric division will lose out to symmetric division. We see a specific example of this in the epidermis (https://www.nature.c...1586-019-1085-7), where asymmetrically division occurs vertically and whilst this supplies new skin cells, it loses out in the long run to symmetrically dividing skin cells, which spread out across the base of the epidermis. This leads to thinner skin with time, as the stem cells that come to dominate the basal layer require ever greater stimulation to supply the epidermis with new cells. If we extend this to the whole body, we find that more quiescent cells will come to dominate the stem cell niches and that this is coincident with a failure of tissue renewal. Interestingly ROCK inhibitors, which cause limited de-differentiation seem to be able to reverse this process in the endothelium (see thread) and the epidermis (in a cyclical manner). This may be because it de-differentiates cells to a progenitor stage, which can then reseed the pool before re-differentiating.

[QuestforLife]

I’m currently doing a continuous cycle of:
• SES (Statin-epitalon/endoluten-Sartan) as previously described for 20 days on, 20 days off
• Senolytics (liposomal vitamin C-azithromycin-doxycycline) for 2/3 days, completed once - report to follow

After various discussions with Turnbuckle I am convinced that increasing stem cell numbers is a good idea, particularly if there really are embryonic like cells still in the body as suggested (https://ahajournals....SAHA.118.314287). This is the key point – these cells have not passed the pluripotency barrier, so can expand without loss of telomeres.

I’ve tried TB’s protocol numerous times, without noticing anything apart from fatigue. Having done my own research, I have my own ideas about how to do this. As I’ve yet to try it, I’m hesitant to post it here. But I’ll include it here as a reminder to myself to keep this thread updated with any positive or negative results. The key thing will be the duration of the stimulating protocol, getting it right may be tricky – we want to wake VSELs (Very Small Embryonic Like stem cells) up, but not differentiate them.

 

VSEL stimulating protocol
Nicotinamide (HDAC inhibition, demethylation)
DHEA or pregnenolone (sex hormones to aid waking up quiescent cells)
VSEL symmetrical expansion
Glycerine Monostearate (mito-fusion)
TMG (for methyl groups)
C60 for stem cell stimulation (to be added as required)

 

During the stimulating protocol it is important not to take TMG or other methyl donors, as the whole idea is to let high dose nicotinamide strip the body of methyl groups. Obviously this would be highly dangerous if done for too long. Based on various papers however, 2 g a day of NAM is safe, even without methyl boosters. But the liver will be using methyl groups up like crazy, hopefully denuding the rest of the body. Limiting methionine and glycerine intake will be helpful during the stimulation phase (although probably not critical)

Further Methodology
The idea is to intentionally deplete body of methyl groups to poise pluripotent cells for differentiation, but then remethylate and expand symmetrically instead.
In more detail: nicotinamide is known as an HDAC inhibitor, probably because it depletes the body of methyl groups, and this is useful in activating long-quiescent stem cells. Simultaneously using increased hormones, (i.e. pregnenolone), may be a further way to wake such cells and poise them for differentiation.
Rather than allowing differentiation, one can then remethylate with TMG whilst simultaneously causing mitochondria to fuse and be stimulated whist in the presence of low ROS. This should increase the chances of symmetrical division.

 

I include for completeness the following, which I posted to TB’s thread:
After doing some reading around the subject, I’m fairy sure you’re wrong Turnbuckle that Threonine is vital to human ESCs (and we’ve all wasted money on buying this expensive supplement – but hey, that’s the life of a self-experimenter). The threonine requirement was shown unequivocally for mouse ESCs (https://www.ncbi.nlm...pubmed/19589965) and then tentatively extended to human ESCs (https://www.frontiersin.org/articles/10.3389/fcell.2014.00018/full ), although they never actually showed it beyond doubt. But since then there have been papers by other authors that have shown beyond doubt that human cells don’t need threonine, instead they use methionine (https://www.cell.com/cell-metabolism/fulltext/S1550-4131(14)00122-3?_returnURL=https%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii%2FS1550413114001223%3Fshowall%3Dtrue ) for the same purpose. This makes sense when you consider a couple of things: stem cells use methylation to remain pluripotent and quiescent; humans don’t have the threonine dehydrogenase enzyme so they can’t use it in the one carbon cycle in mitochondria to make methyl groups. It is not exactly clear why we can’t use glycine in the absence of methionine but it might be our limited supply is used for other things.

[QuestforLife]

Some more ideas on aging and skin aging in particular: Stem cells are activated by stressors. Over a lifetime it makes sense that the more easily activated stem cells (and there is a range of responsiveness in all cells) will be depleted first – because asymmetric division will lose out to symmetric division. We see a specific example of this in the epidermis (https://www.nature.c...1586-019-1085-7), where asymmetrically division occurs vertically and whilst this supplies new skin cells, it loses out in the long run to symmetrically dividing skin cells, which spread out across the base of the epidermis. This leads to thinner skin with time, as the stem cells that come to dominate the basal layer require ever greater stimulation to supply the epidermis with new cells. If we extend this to the whole body, we find that more quiescent cells will come to dominate the stem cell niches and that this is coincident with a failure of tissue renewal. Interestingly ROCK inhibitors, which cause limited de-differentiation seem to be able to reverse this process in the endothelium (see thread) and the epidermis (in a cyclical manner). This may be because it de-differentiates cells to a progenitor stage, which can then reseed the pool before re-differentiating.

 

Looking at 'Stem cell competition orchestrates skin homeostasis and ageing', I think their conclusions require further elaboration. Here is their abstract:

 

 

Stem cells underlie tissue homeostasis, but their dynamics during ageing—and the relevance of these dynamics to organ ageing—remain unknown. Here we report that the expression of the hemidesmosome component collagen XVII (COL17A1) by epidermal stem cells fluctuates physiologically through genomic/oxidative stress-induced proteolysis, and that the resulting differential expression of COL17A1 in individual stem cells generates a driving force for cell competition. In vivo clonal analysis in mice and in vitro 3D modelling show that clones that express high levels of COL17A1, which divide symmetrically, outcompete and eliminate adjacent stressed clones that express low levels of COL17A1, which divide asymmetrically. Stem cells with higher potential or quality are thus selected for homeostasis, but their eventual loss of COL17A1 limits their competition, thereby causing ageing. The resultant hemidesmosome fragility and stem cell delamination deplete adjacent melanocytes and fibroblasts to promote skin ageing. Conversely, the forced maintenance of COL17A1 rescues skin organ ageing, thereby indicating potential angles for anti-ageing therapeutic intervention.

What is actually happening, as I stated in my last post on the subject, is that stress on the skin is selecting a population of more stress resistant stem cells (as defined by their expression of COL17A1). This leads to less asymmetric division and skin replacement and therefore thinner skin, which presumably then leads to higher stress on the remaining cells and the process continues. Eventually you are left with a population of super stress resistant basal stem cells, analogous to the super-quiescent VSELs that exist in other tissues. They authors say ‘eventual loss of COL17A1 limits their competition, thereby causing ageing’, but actually this is only the final part of skin aging. Overly stress resistant cells is as much a part of the problem as loss of the cells that are more easily stimulated.

They leave the best for last, showing that low doses of a ROCK inhibitor (or high doses of an anti-oxidant, Apocynin) both increase wound healing in mice, and prevent the thinning of skin in a model of aged skin.
My research has shown that statins are effective ROCK inhibitors even at very low doses, and have even been used in increase wound healing (https://www.ncbi.nlm...pubmed/28740761).

How is this relevant to my skin rejuvenation approach?
I am 40 and never noticed any aging at 35. But by my late 30s, and coinciding with a prolonged period of extreme stress, when I took little care of myself, my skin dried out and non-permanent or semi-permanent wrinkles on my forehead deepened and become permanent. A number of finer lines also appeared around my eyes and also around the deeper lines of my forehead. Various skin creams have since managed to re-hydrate my skin and stop any further deterioration, plus achieved a reduction of the newer, finer lines. But the deeper lines have barely changed.
After various experiments, I now think that I can achieve thicker skin with a simvastatin skin cream. But in order to genuinely reverse deeper lines, I now believe I will need to take more drastic action.  For this I will use derma-needling, followed by my statin skin cream, enabling deeper penetration (https://www.ncbi.nlm...pubmed/27530764) and better healing. In fact, my statin cream should allow more regular derma-needling, faster and better healing, and therefore a quicker improvement in appearance. To boost collagen production I will also orally take Lipsomal Vit C and Glycine. This paper (https://www.ncbi.nlm.nih.gov/pubmed/?term=High+glycine+concentration+increases+collagen+synthesis+by%C2%A0articular+chondrocytes+in%C2%A0vitro%3A+acute+glycine+deficiency+could+be+an%C2%A0important+cause+of%C2%A0osteoarthritis) shows why glycine is the best amino acid to take to boost collagen production, with no downside (collagen production at higher doses does not invert, unlike with Lysine). See attached pic.

 

Attached Thumbnails

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Edited by QuestforLife, 26 September 2019 - 12:19 PM.

[Kentavr]

7 months after my last methylation DNAage test I got another one.

I deliberately took the samples just after I had completed the last statin-sartan course March, and there has been an improvement, albeit a small one.

Apparently I am now only 38 epigenetically, 2.4 years less than my actual 40 years.

My previous result, 7 months prior, said I was only 0.7 years less than my chronological age at that time (39).

So some tentative evidence for my protocol being effective.

Thanks for the comment.

I periodically use metformin (1000 mg glucophage long) + sartans (telmisartan 20 mg).

I have rosuvastatin 10 mg, but I have not used it yet, since statins damage mitochondria.

What drugs do you use and at what dosage?

[QuestforLife]

Thanks for the comment.

I periodically use metformin (1000 mg glucophage long) + sartans (telmisartan 20 mg).

I have rosuvastatin 10 mg, but I have not used it yet, since statins damage mitochondria.

What drugs do you use and at what dosage?

It's all in this thread. I use 5mg atorvastatin, 12.5mg losartan with a telomerase activator like epitalon or endoluten for a 20 day cycle. The low dose and short cycle means I benefit from the ROCK inhibition, which generates progenitor cells, but not from any side effects such as chloresterol lowering (or only very weakly).

Actually Kentavr, it was you who first gave me the idea for this protocol,from a paper by Janic et al on using a low dose statin and sartan to rejuvenate the endothelial lining of the arteries.

I've also been experimenting with a statin skin cream. Interestingly a cycle seems to work best there too - so we periodically should be building up progenitor/stem cells, but then we need to let them differentiate to get the maximum benefits.